AIM: To study the toxicity of genipin-a kind of geniposide metabolites induced human tubular epithelial cells HK-2 and its effect on NLRP3 pathway. METHODS: The dose of GP on HK-2 cells were preliminarily determined by CCK8 method, the apoptosis or necrosis rate of HK-2 cells was detected by Hoechst 33342/PI, the level of LDH release and reactive oxygen species was detected by Kits, and mitochondrial membrane potential and intracellular calcium ion concentration were detected by high content imaging. Real-time PCR detected mRNA levels of kindey injury factor-1, osteopontin, NLRP3, Caspase-1, interleukin 1β, and interleukin 18. RESULTS: Compared with the 0 μg/mL group, GP>50 μg/mL significantly reduced cell viability (P<0.05, P<0.01), and the IC50 value was 110.50 μg/mL. Set the control group, the low, medium and high dose groups of GP (50, 100, 200 μg/mL); Compared with the control group, the cell density decreased in the medium and high dose groups of GP, and the PI positivity, LDH release, ROS, Ca2+ concentration increased significantly, MMP decreased significantly, and KIM-1, OPN, NLRP3, Caspase-1, IL-1β and IL-18mRNA levels increased significantly (P<0.05, P<0.01). CONCLUSION: GP may activate NLRP3 by increasing ROS and Ca2+, decreasing MMP and causing HK-2 cell damage.